Abstract: Approximately 2 billion people worldwide are infected with helminths and the resulting helminthiasis is a heavy health burden for developing countries. Parasitic helminths are a class of multicellular parasites, mainly including trematodes, tapeworms and nematodes, wtih complex life cycle involving multiple developmental stages and typically one or more hosts. Understanding the growth, development, pathogenesis and transmission of these parasites at the molecular level is of great significance for the diagnosis and treatment of helminthiasis. Genetic manipulations, which alter the expression level of target genes, have greatly promoted the biomedical research. In recent years, with the release of genomic data of worms, genetic manipulation techniques, such as RNA interference and clustered regularly interspaced short palindromic repeats (CRISPR) gene editing, have been increasingly applied in the studies of parasitic helminths. This article reviews the progress of genetic manipulations in important medical worms, as well as the methods of genetic manipulations, which would be expected to inspire the future functional study in parasitic helminths.

Objective To investigate the role of protein kinase C (PKC) in mechanical ventilation-induced lung injury in rats.Methods Thirty healthy male Wistar rats,weighing 250-300 g,were randomly divided into 5 groups (n =6 each):control group (group C),small tidal volume group (group S),small tidal volume and PKC inhibitor group (group S + P),large tidal volume group (group L),and large tidal volume and PKC inhibitor group (group L + P).VT =42 ml/kg,RR =40 bpm,I∶E =1∶ 2,PEEP =0,FiO2 =21％ in groups L and L + P,while VT=7 ml/kg,RR=40 bpm,I∶E=1∶2,PEEP=0,FiO2 =21％ in groups S and S+P.The rats were only tracheostonized in group C,while the rats were mechanically ventilated for 4 h in the other four groups.PKC inhibitor bisindolylmaleinide Ⅰ 0.12 mg/kg was injected intramuscularly 1 h before anesthesia in groups S + P and L + P.The animals were sacrificed immediacy after tracheotomy in group C,and at 4 h of ventilation in the other four groups and lungs were removed for calculation of wet/dry lung weight ratio (W/D ratio) and for microscopic examination.The expression of occludin was determined in the lung tissues by Western blot.Results Compared with group C,W/D ratio was significantly increased and the expression of occludin was down-regulated in the other four groups (P ＜ 0.05).Compared with group S,W/D ratio was significantly increased and the expression of occludin was down-regulated in group L,and W/D ratio was decreased and the expression of occludin was up-regulated in group S + P (P ＜ 0.01).W/D ratio was significantly lower and the expression of occludin was higher in group L + P than in group L (P ＜ 0.01).The pathological changes were attenuated in groups S + P and L + P as compared with groups S and L.Conclusion PKC is involved in mechanical ventilation-induced lung injury in rats.

Objective To investigate the prevalence of gastroeaophageat reflux disease(GERD)in the end stage renal disease patients(ESRD)with maintenance hemodialysis and the pathogenetic factors of GERD.Methods 60 cases who had heart burn and acid regurgitation symptoms were enrolled,31 cases of them were ESRD patients with maintenance hemodialysis,29 patients who were renal diseases free as control group.GERD was diagnosed by three methods:endoscopy,24h esophageal pH monitoring,endoscopic esophageal biopsy.GERD was established if any of these tests was positive.HP infection and serum gastrin were assessed too.Results (1)The prevalences of GERD in two groups were 83.9%,79.3% respectively and had no significant difference(P＞0.05).(2)The prevalence of HP infection in the ESRD group was significantly lower than control group(19 4% vs 51.7%,P＜0.05).(3)The concentration of serum gastrin in the ESRD group and control group was(102.61±30.80)ng/ml and (83.41±20.61)ng/ml,two group had significant difference(P＜0.05).Conclusion The prevalence of GERD inthe ESRD patients was higher than expected,and was correlated with absence of HP infection and hypergastinemia.

Objective To evaluate the role of Src tyrosine kinase in damage to alveolar epithelial cells caused by mechanical stretch.Methods MLE-12 cells cultured in vitro were randomly divided into 3 groups using a random number table:mechanical stretch group (group S),dimethyl sulfoxide control group (group D),and Src tyrosine kinase inhibitor PP2 group (group P).In D and P groups,dimethyl sulfoxide 30 μl/ml and PP2 100 μmol/L were added to the culture medium,respectively,and the cells were then cultured for 30 min.The cells underwent mechanical stretch for 8 h with frequency of0.5 Hz and amplitude of 20％ in the three groups.At 0,2,4 and 8 h of mechanical stretch,MLE-12 cells in 3 wells of each group were collected for determination of cell apoptosis with flow cytometry and expression of occludin using Western blot.The apoptosis rate was calculated.Results Compared with S group,no significant changes were found in the apoptosis rate and expression of occludin at each time point in group D,and the apoptosis rate was significantly decreased,and the expression of occludin was up-regulated at 2,4 and 8 h of mechanical stretch in group P.Conclusion The activation of Src tyrosine kinase is involved in damage to alveolar epithelial cells caused by mechanical stretch.

Objective To evaluate the effect of recombinant human annexin A5 on the expression of phosphorylated protein kinase C alpha (p-PKCα) and p120-catenin during endotoxin-induced damage to cardiomyocytes.Methods H9c2 cells cultured in vitro were randomly divided into 3 groups (n=18 each) using a random number table:control group (group C),endotoxin group (group L),and recombinant human annexin A5 group (group A).Recombinant human annexin A5 (final concentration 5 ng/ml) was added,and the cells were incubated for 2 h in group A,and then lipopolysaccharide (final concentration 1 μg/ml) was added,and the cells were incubated for 4 h in L and A groups.At 4 h of incubation,cell apoptosis was detected using the cell apoptosis detection kit,the intercellular space was measured using the confocal microscopy,and the expression of p-PKCα and p120-catenin was determined by Western blot.The apoptosis rate was calculated.Results Compared with group C,the apoptosis rate was significantly increased,the intercellular space was significantly widened,the expression of p120-catenin was significantly down-regulated,and the expression of p-PKCα was significantly up-regulated in group L (P＜0.05).Compared with group L,the apoptosis rate and intercellular space were significantly decreased,the expression of p120-catenin was significantly up-regulated,and the expression of p-PKCα was significantly down-regulated in group A (P＜0.05).Conclusion Recombinant human annexin A5 can inhibit phosphorylation of PKCα and up-regulate the expression of p120-catenin,thus attenuating endotoxin-induced damage to cardiomyocytes.

Objective Although principal components analysis profiles greatly facilitate the visualization and interpretation of the multivariate data,the quantitative concepts in both scores plot and loading plot are rather obscure.This article introduced three profiles that assisted the better understanding of metabolomic data.Methods The discriminatory profile,heat map,and statistic profile were developed to visualize the multivariate data obtained from high-throughput GC-TOF-MS analysis.Results The discriminatory profile and heat map obviously showed the discriminatory metabolites between the two groups,while the statistic profile showed the potential markers of statistic significance.Conclusion The three types of profiles greatly facilitate our understanding of the metabolomic data and the identification of the potential markers.

Objective: To investigate the association between dietary selenium intake and hypertension among Zhejiang residents . Methods: By multistage stratified random sampling method,four urban sites and two rural sites out of Zhejiang Province,four communities or villages out of each site,then 20 households out of each community or village were selected,and all the family members of the selected households were recruited as participants. The questionnaire of Chinese Health and Nutrition Survey was used to collect information about socio-demographic characteristics and dietary selenium intake. The blood pressure,blood lipid and other data were collected via physical examination. A multivariate logistic regression model was used to analyze the relationship between dietary selenium intake and hypertension . Results: A total of 1 222 participants with complete dietary selenium intake data were included for analysis. The number（%）of participants with selenium intake higher than the level of estimated average requirement（EAR）,between the levels of EAR and recommanded nutrient intake （RNI）,between the levels of RNI and upper intake（UI）and higher than the level of UI were 729 （59.66%）,151（12.36%）,341（27.91%）and 1（0.01%）,respectively. There were 283 （30.53%）patients with hypertension out of 927 participants examined. The mean amount of selenium intake in patients with hypertention was（43.06±20.96）μg/d,which was significantly lower than（51.56±30.06） μg/d in non-hypertention participants（P<0.05）. After adjusting for age,body mass index,total cholesterol,triglyceride and diabetes mellitus in the multivariate logistic regression model,dietary selenium intake significantly reduced the risk of hypertension（OR=0.985,95%CI：0.978-0.993） . Conclusion About 60% of residents in Zhejiang Province had lower dietary selenium intake than estimated average requirement. Higher selenium intake was associated with lower risk of hypertension.

Objective:In general, patients with seropositive rheumatoid arthritis (RA) are considered to show an aggressive disease course. However, the relationship between the two subgroups in disease severity is controversial. Our study is aimed to compare the clinical characteristics and prognosis of double-seropositive and seronegative RA in China through a real-world large scale study.Methods:RA patients who met the 1987 American College of Rheumatology (ACR) classification criteria or the 2010 ACR/European Anti-Rheumatism Alliance RA classification criteria, and who attended the 10 hospitals across the country from September 2015 to January 2020, were enrolled. According to the serological status, patients were divided into 4 subgroups [rheumatoid factor (RF)(-) anti-cyclic citrullinated peptide (CCP) antibody (-), RF(+), RF(+) anti-CCP antibody(+), anti-CCP antibody(+)] and compared the disease characteristics and treatment response. One-way analysis of variance was used for measurement data that conformed to normal distribution, Kruskal-Wallis H test was used for measurement data that did not conform to normal distribution; paired t test was used for comparison before and after treatment within the group if the data was normally distributed else paired rank sum test was used; χ2 test was used for count data. Results:① A total of 2 461 patients were included, including 1 813 RF(+) anti-CCP antibody(+) patients (73.67%), 129 RF(+) patients (5.24%), 245 RF(-) anti-CCP antibody(-) patients (9.96%), 74 anti-CCP antibody(+) patients (11.13%). ② Regardless of the CCP status, RF(+) patients had an early age of onset [RF(-) anti-CCP antibody(-) (51±14) years old, anti-CCP antibody(+) (50±15) years old, RF(+) anti-CCP antibody(+) (48±14) years old, RF(+)(48±13) years old, F=3.003, P=0.029], longer disease duration [RF(-) anti-CCP antibody(-) 50 (20, 126) months, anti-CCP antibody(+) 60(24, 150) months, RF(+) anti-CCP antibody(+) 89(35, 179) months, RF(+) 83(25, 160) months, H=22.001, P<0.01], more joint swelling counts (SJC) [RF(-) anti-CCP antibody(-) 2(0, 6), Anti-CCP antibody(+) 2(0, 5), RF(+) anti-CCP antibody(+) 2(0, 7), RF(+) 2(0, 6), H=8.939, P=0.03] and tender joint counts (TJC) [RF(-) anti-CCP antibody(-) 3(0, 8), anti-CCP antibody(+) 2(0, 6), RF(+) anti-CCP antibody(+) 3(1, 9), RF(+) 2(0, 8), H=11.341, P=0.01] and the morning stiff time was longer [RF(-) anti-CCP antibody(-) 30(0, 60) min, anti-CCP antibody(+) 20(0, 60) min, RF(+) anti-CCP antibody(+) 30(10, 60) min, RF(+) 30(10, 60) min, H=13.32, P<0.01]; ESR [RF(-) anti-CCP antibody(-) 17(9, 38) mm/1 h, anti-CCP antibody(+) 20(10, 35) mm/1 h, RF(+) anti-CCP antibody(+) 26(14, 45) mm/1 h, RF(+) 28(14, 50) mm/1 h, H=37.084, P<0.01] and CRP [RF(-) anti-CCP antibody(-) 2.3 (0.8, 15.9) mm/L, Anti-CCP antibody(+) 2.7(0.7, 12.1) mm/L, RF(+) anti-CCP antibody(+) 5.2(1.3, 17.2) mm/L, RF (+) 5.2(0.9, 16.2) mm/L, H=22.141, P<0.01] of the RF(+)patients were significantly higher than RF(-) patients, and RF(+) patients had higher disease severity(DAS28-ESR) [RF(-) anti-CCP antibody(-) (4.0±1.8), anti-CCP antibody(+) (3.8±1.6), RF(+) anti-CCP antibody(+) (4.3±1.8), RF(+) (4.1±1.7), F=7.269, P<0.01]. ③ The RF(+) anti-CCP antibody(+) patients were divided into 4 subgroups, and it was found that RF-H anti-CCP antibody-L patients had higher disease severity [RF-H anti-CCP antibody-H 4.3(2.9, 5.6), RF-L anti-CCP antibody-L 4.5(3.0, 5.7), RF-H anti-CCP antibody-L 4.9(3.1, 6.2), RF-L anti-CCP antibody-H 2.8(1.8, 3.9), H=20.374, P<0.01]. ④ After 3-month follow up, the clinical characteristics of the four groups were improved, but there was no significant difference in the improvement of the four groups, indicating that the RF and anti-CCP antibody status did not affect the remission within 3 months. Conclusion:Among RA patients, the disease activity of RA patients is closely related to RF and the RF(+) patients have more severe disease than RF(-) patients. Patients with higher RF titer also have more severe disease than that of patients with low RF titer. After 3 months of medication treatment, the antibody status does not affect the disease remission rate.

Multiple myeloma (MM) is still an incurable hematologic malignancy, which is eagerly to the discovery of novel therapeutic targets and methods. N-acetyltransferase 10 (NAT10) is the first reported regulator of mRNA acetylation that is activated in many cancers. However, the function of NAT10 in MM remains unclear. We found significant upregulation of NAT10 in MM patients compared to normal plasma cells, which was also highly correlated with MM poor outcome. Further enforced NAT10 expression promoted MM growth in vitro and in vivo, while knockdown of NAT10 reversed those effects. The correlation analysis of acetylated RNA immunoprecipitation sequencing (acRIP-seq) and ribosome profiling sequencing (Ribo-seq) combined with RIP-PCR tests identified centrosomal protein 170 (CEP170) as an important downstream target of NAT10. Interfering CEP170 expression in NAT10-OE cells attenuated the acceleration of cellular growth caused by elevated NAT10. Moreover, CEP170 overexpression promoted cellular proliferation and chromosomal instability (CIN) in MM. Intriguingly, remodelin, a selective NAT10 inhibitor, suppressed MM cellular growth, induced cellular apoptosis in vitro and prolonged the survival of 5TMM3VT mice in vivo. Collectively, our data indicate that NAT10 acetylates CEP1 70 mRNA to enhance CEP170 translation efficiency, which suggests that NAT10 may serve as a promising therapeutic target in MM.